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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry
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Fluorescence intensity normalisation: correcting for time effects in large-scale flow cytometric analysis.

Calliope A Dendrou1, Erik Fung, Laura Esposito

  • 1Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.

Advances in Bioinformatics
|January 6, 2010
PubMed
Summary
This summary is machine-generated.

Normalizing flow cytometry data with beads or alternative methods improves repeatability for genetic association studies. This enhances the analysis of protein levels in immune cells over extended experimental periods.

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Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction

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Area of Science:

  • Immunology
  • Genetics
  • Biotechnology

Background:

  • Genome-wide association studies (GWAS) require linking molecular traits to disease alleles.
  • Gene expression quantitative trait loci (eQTL) link mRNA levels to genetic variations.
  • Flow cytometry offers valuable protein-level data from multiple cell subsets, but faces throughput limitations.

Purpose of the Study:

  • To develop normalization methods for flow cytometry data to enable genetic association studies.
  • To improve the repeatability of protein phenotype measurements over extended experimental timelines.
  • To assess the utility of normalising fluorospheres and propose bead-free alternatives.

Main Methods:

  • Investigated the use of normalising fluorospheres (broad spectrum and spectrum matched) for flow cytometry.
  • Assessed the repeatability of CD25-APC mean fluorescence intensity on CD4(+) memory T cells.
  • Developed and evaluated two alternative normalization procedures without beads.

Main Results:

  • Normalising fluorospheres significantly improved the repeatability of flow cytometry measurements.
  • Spectrum-matched beads showed potential for enhanced normalization.
  • Alternative bead-free normalization methods were proposed and assessed.

Conclusions:

  • Normalization is crucial for comparing flow cytometry data collected over extended periods.
  • Normalising fluorospheres enhance the reliability of protein-level phenotypic data for genetic studies.
  • Bead-free normalization strategies offer viable alternatives for genetic association studies using flow cytometry.